A simple method for detection of human factor IX inhibitor using ELISA.
We developed a simple ELISA for FIX inhibitors (FIXinh). The relationship between absorbance and various dilutions of plasma containing known FIXinh (28 Bethesda units/ml) was linear, ranging from 200 to 25,600-fold dilution on semi-log paper. Converted into the value of Bethesda Inhibitor Assay, ELISA can detect as low as 0.219 Bethesda units/ml. Absorbance for 30 normal individuals was 0.213 +/- 0.008, which corresponds to < 0.44 Bethesda units/ml. With ELISA, only the value for hemophilia B with FIXinh, which was revealed as positive by Bethesda Inhibitor Assay, was significantly higher than that of normal controls, hemophilia B without FIXinh, hemophilia A with FVIIIinh and systematic lupus erythematosus (SLE) with lupus anticoagulant (LA). The value in the patient with SLE, who had a high level of FVIIIinh (1000 Bethesda units/ml), was 3 Bethesda units/ml of FIXinh with Bethesda Inhibitor Assay, which was not significantly higher than that of normal subjects with ELISA. FIXinh in hemophilia B patients by ELISA for FIXinh was not affected by the presence of other anticoagulants.
Takamiya
O
O
Department of Medical Technology, School of Allied Medical Sciences, Shinshu University, Asahi, Japan.
Kinoshita
S
S
D000328:Adult; D002118:Calcium / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005164:Factor IX / Q000037:antagonists & inhibitors* / Q000276:immunology*; D005260:Female; D006467:Hemophilia A / Q000276:immunology; D002836:Hemophilia B / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000145:classification; D007518:Isoantibodies / Q000097:blood*; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008297:Male; D008875:Middle Aged
D007074:Immunoglobulin G; D007518:Isoantibodies; D005164:Factor IX; D002118:Calcium